A defined bacterial culture protective against Salmonella typhimurium cecal colonization in broiler chicks was derived utilizing a continuous-flow (CF) culture apparatus. Chicks receiving the CF culture in combination with a diet containing dietary lactose were protected against cecal colonization by S. typhimurium. The culture consisted of a mixture of gram-positive and gram-negative facultative and strictly anaerobic bacteria. The isolates were identified as Enterococcus avium, two strains of Enterococcus faecalis (designated A and B), Lactococcus lactis, Lactobacillus animalis, a Lactobacillus that could not be identified to species level (designated strain CMS), Citrobacter freundii Escherichia coli, E. fergusonii, Bifidobacterium animalis, and Propionibacterium acidipropionici. Results indicated that CF cultures can be used as a tool to identify bacteria which are antagonistic to S. typbimurium in the chick cecum.